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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

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qKAT: Quantitative Semi-automated Typing of Killer-cell Immunoglobulin-like Receptor Genes
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qKAT: Quantitative Semi-automated Typing of Killer-cell Immunoglobulin-like Receptor Genes

Published on: March 6, 2019

Quenched autoligation probes.

Adam P Silverman1, Hiroshi Abe, Eric T Kool

  • 1Department of Chemistry, Stanford University, Stanford, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 13, 2008
PubMed
Summary
This summary is machine-generated.

Quenched autoligation (QUAL) probes offer a single-step method for detecting DNA and RNA. These fluorescent probes can identify minute nucleic acid variations in various sample types, including living cells.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Analytical Chemistry

Background:

  • Oligonucleotide-based detection methods are crucial for molecular diagnostics.
  • Existing fluorescent probes often require multiple steps or probe removal.
  • There is a need for sensitive and specific nucleic acid detection with simplified workflows.

Purpose of the Study:

  • To describe the preparation and application of quenched autoligation (QUAL) probes.
  • To demonstrate the utility of QUAL probes for sensitive nucleic acid detection.
  • To highlight the single-step, no-wash signal generation of QUAL probes.

Main Methods:

  • Synthesis of dabsyl quencher linker and QUAL probes using DNA synthesizer.
  • Application of QUAL probes for detecting nucleic acids in solution.
  • Utilizing QUAL probes for detecting RNA in bacterial and human cells.

Main Results:

  • QUAL probes provide a "lighting up" fluorescent signal in a single step.
  • The probes can detect DNA and RNA in solution, on surfaces, and within cells.
  • QUAL probes exhibit high specificity, distinguishing single nucleotide differences in targets.

Conclusions:

  • QUAL probes represent an efficient and versatile tool for nucleic acid detection.
  • The single-step, no-wash nature simplifies detection protocols.
  • These probes have broad applications in biological research and diagnostics.